Methods of evaluating specific cellular functions of receptor protein tyrosine kinases in a ligand independent manner

Abstract

The invention relates to methods of evaluating the specific function of a receptor protein tyrosine kinase in cells. The methods activate the receptor in a ligand independent fashion. In addition, the invention includes methods of identifying compounds that modulate receptor protein tyrosine kinase function.

What is claimed is:  
     
         1 . A method of evaluating a function of a receptor protein tyrosine kinase comprising the following steps: 
 (a) transfecting a nucleic acid vector into cells, wherein said vector encodes a chimera comprising an extracellular region and an intracellular region, wherein said intracellular region is from said receptor protein tyrosine kinase;    (b) contacting said cells with an antibody, wherein said antibody has specific binding affinity to said extracellular region; and    (c) monitoring an effect on said cells.    
     
     
         2 . The method of  claim 1 , where said extracellular region consists of an extracellular region of a receptor protein tyrosine kinase selected from the group consisting of TRK, EGFR, PDGFR, and RET.  
     
     
         3 . The method of  claim 1 , wherein said intracellular region consists of an intracellular region of an orphan receptor protein tyrosine kinase.  
     
     
         4 . The method of  claim 3 , wherein said orphan receptor protein tyrosine kinase is selected from the group consisting of C-RET, SEK, MCK-10, AXL, TYRO3, MER, EPH, ECK, EEK, ERK, ELK, EHK1, EHK2, SEK, HEK, HEK2, MYK1, CEK9, MYK2, MDK1, IRR, CCK4, RYK, DDR, TYRO10, ROS, LTK, ALK, ROR1, ROR2, and TOR.  
     
     
         5 . The method of  claim 4 , wherein said orphan receptor protein tyrosine kinase is C-RET.  
     
     
         6 . The method of  claim 1 , wherein said cells and said extracellular region are from different species.  
     
     
         7 . The method of  claim 6 , wherein said cells are mammalian.  
     
     
         8 . The method of  claim 6 , wherein said extracellular region is isolated from a chicken.  
     
     
         9 . The method of  claim 1 , wherein said antibody has specific binding affinity to a TRK extracellular region isolated from a chicken.  
     
     
         10 . The method of  claim 1 , wherein said effect is a change or an absence of a change in cell phenotype.  
     
     
         11 . A method of identifying compounds that modulate the function of a receptor protein tyrosine kinase in cells, wherein said method comprises the following steps: 
 (a) transfecting a nucleic acid vector into said cells, wherein said vector encodes a chimera comprising an extracellular region and an intracellular region, wherein said intracellular region is from said receptor protein tyrosine kinase;    (b) contacting said cells with one or more compounds;    (c) contacting said cells with an antibody, wherein said antibody has specific binding affinity to said extracellular region; and    (d) monitoring an effect on said cells.    
     
     
         12 . The method of  claim 11 , wherein said extracellular region consists of an extracellular region of a receptor protein tyrosine kinase selected from the group consisting of TRK, EGFR, PDGFR, and RET.  
     
     
         13 . The method of  claim 11 , wherein said intracellular region consists of intracellular region of an orphan receptor protein tyrosine kinase.  
     
     
         14 . The method of  claim 13 , wherein said orphan receptor protein tyrosine kinase is selected from the group consisting of C-RET, SEK, MCK-10, AXL, TYRO3, MER, EPH, ECK, EEK, ERK, ELK, EHK1, EHK2, SEK, HEK, HEK2, MYK1, CEK9, MYK2, MDK1, IRR, CCK4, RYK, DDR, TYRO10, ROS, LTK, ALK, ROR1, ROR2, and TOR.  
     
     
         15 . The method of  claim 14 , wherein said orphan receptor protein tyrosine kinase is C-RET.  
     
     
         16 . The method of  claim 11 , wherein said cells and said extracellular region are from different species.  
     
     
         17 . The method of  claim 16 , wherein said cells are mammalian.  
     
     
         18 . The method of  claim 16 , wherein said extracellular region is isolated from a chicken.  
     
     
         19 . The method of  claim 11 , wherein said antibody has specific binding affinity to a TRK extracellular region isolated from a chicken.  
     
     
         20 . The method of  claim 11 , wherein said effect is a change or an absence of a change in cell phenotype.  
     
     
         21 . The method of  claim 11 , wherein said effect is a change or an absence of a change in the catalytic activity of said intracellular region.  
     
     
         22 . The method of  claim 11 , wherein said effect is a change or an absence of a change in an interaction between said intracellular region and a natural binding partner.  
     
     
         23 . A method of identifying compounds that modulate the function of C-RET receptor protein tyrosine kinase comprising the following steps: 
 (a) expressing said C-RET in cells;    (b) contacting said cells with one or more compounds; and    (c) monitoring an effect on said cells.    
     
     
         24 . The method of  claim 23 , wherein said effect is a change or an absence of a change in cell phenotype.  
     
     
         25 . The method of  claim 23 , wherein said effect is a change or an absence of a change in catalytic activity of said C-RET receptor.  
     
     
         26 . The method of  claim 23 , wherein said effect is a change or an absence of a change in the interaction between said C-RET receptor and a natural binding partner.